Blood mRNA detection kit and detection method

Abstract

The invention discloses a blood mRNA detection kit and a detection method. The blood mRNA detection kit contains nine pairs of specific multiple PCR primers shown in SEQ ID NO.1-18 as shown in description and 16 SNP single-base extension primers shown in SEQ ID NO.19-34 as shown in description, and is used for detecting 16 SNP molecular markers on five kinds of blood specific mRNA molecules, operation of detecting blood RNA, which accounts for as low as 1% or of which the content is as low as 0.1 ng, in complex mixed stains can be completed, blood can be separated from other body fluid and skin tissues precisely, a source is traced back to fast and accurately, and a suspect is positioned.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a blood detection kit and a detection method. Background technique [0002] In forensic work, complex mixed plaque is one of the most common biological samples, and it has always been the focus and difficulty of inspection. Complex mixed plaques include: (1) mixed plaques composed of multiple body fluids; (2) extremely unbalanced mixed plaques (where the composition ratio of one component is less than 10%). The mainstream technology for analyzing mixed plaques composed of multiple body fluids at home and abroad is to type STR based on single cell separation technology, capillary electrophoresis or next-generation sequencing platforms. [0003] Some mRNA molecules in blood are specific to body fluids and tissues. These specific mRNAs can only be expressed and detected in blood, while other body fluids do not. At present, in forensic medicine, specific mRNA molecules...

AI Technical Summary

Problems solved by technology

At present, in forensic science, specific mRNA molecules cannot be used alone to identify individuals, and are only used to infer the type of body fluid spots, and to use mRNA and DNA co-extraction technology to achieve spot identification and individual identification

However, although the combination of mRNA molecules and DNA has achieved the purpose of identifying the type of body fluid spots and individual recognition, it has not solved the problem of complex mixed spots.

[0004] Although the next-generation sequencing platform has a good ability to analyze simple mixed spots, it has insurmountable obstacles in the analysis of complex mixed spots in terms of sequencing fragment splicing, sequencing errors, and the detection of small components of unbalanced mixed spots.

In the traditional analysis method, events such as the loss of alleles, the appearance of new alleles, and shadow bands will also occur. During the analysis, multiple individuals will share the same allele, which makes it difficult to interpret the identification results, and subjective deviations even occur. mistake

For mixed plaques with an extremely unbalanced composition ratio, it is difficult to find the cell components of secondary donors (<10%), resulting in the inability to break through the bottleneck of detection sensitivity for complex mixed plaques (5~10%)

[0005] Moreover, traditional methods can only use the analysis results in mixed spots to compare with suspects. Therefore, it is difficult to directly identify suspects using blood information in mixed spots, which is a major problem in the current research on complex mixed spots.

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